An Introductory Guide to Light Microscopy PDF
Preview An Introductory Guide to Light Microscopy
An Introductory Guide to Light Microscopy 16 Apr to 14 May 2007 Michael Hooker Microscopy Facility Michael Chua Wendy Salmon An Introductory Guide to Light Microscopy - Five Talk Plan • Apr 16. A brief perspective of microscopy, theory of operation, key parts of a typical microscope for transmitted light, Kohler illumination, the condenser, objectives, Nomarski, phase contrast, resolution • Apr 23. Fluorescence: Why use it, fluorescence principals, contrast, resolution, filters, dichroic filter cubes, immuno staining, fluorescent proteins, dyes. • Apr 30. Detectors, sampling & digital images: Solid state digital cameras, Photomultipliers, noise, image acquisition, Nyquist criterion/resolution, pixel depth, digital image types/color/compression • May 07. Confocal Microscopy: Theory, sensitivity, pinhole, filters, 3-D projection/volume renders • May 14. Advanced Fluorescence/Confocal: Live cell imaging, co- localization, bleed through/cross talk, FRAP, fluorescence recovery after photobleaching, deconvolution Today’s Plan • Brief perspective / evolution of imaging • Basic Optics • Parts of a microscope & Kohler Illumination • Objectives • Modes of Imaging common in Biomedical Research History and Evolution • 4 Centuries of light microscopy! Zeiss compound microscope with confocal scan head (2001) ~400 years Janssen Zacharias Janssen was a Dutch spectacle-maker credited with inventing the first compound microscope in ~1590. Magnifications ~3x to ~9x. Huygens Christiaan Huygens (1629–1695) was a Dutch mathematician, astronomer and physicist developed an improved two lens eye piece. Optical errors in two half curved lenses tend to cancel out. Can get more magnification. Hooke Robert Hooke improved the design of the new compound microscope, including a light source ~1655. Developed the micrometer. Resolution ~5 um. Leeuwenhoek Anton van Leeuwenhoek simple microscope (~1675) used a single lens which yielded high magnifications (~70x to ~300x) and excellent resolution (~1 um). He reported seeing many kinds of microorganisms including bacteria! Chester Moore Hall & John Dollond • Achromatic refracting lens was invented in 1733 by an English barrister named Chester Moore Hall • Patented by John Dollond. Lister Tight focus Spherical aberration Joseph Jackson Lister (1786-1869) design and construct superior complex lenses by combining lenses of crown and flint glasses of different dispersion, but separated in order to both correct chromatic aberration and minimize spherical aberration. Abbe Ernst Abbe (1840-1905) applied mathematical principles to the design of lenses, which dramatically facilitated the manufacturing high quality optical instruments by the Carl Zeiss corporation. Abbe's realization that the performance of a microscope was limited by the diffraction of light was not well accepted for decades. The Abbe limit formula
The list of books you might like
